Crystalline and salt forms of ppar agonist compounds

ABSTRACT

This disclosure relates to salt forms of compounds capable of activating PPARδ for use in drug substance and drug product development, and related compositions and methods.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/404,474, filed on Oct. 5, 2016. The entire teachings of theaforementioned application are incorporated herein by reference.

FIELD OF THE INVENTION

This disclosure relates to solid forms of compounds capable ofactivating PPARδ for use in drug substance and drug product development,and related compositions and methods.

BACKGROUND OF THE INVENTION

Peroxisome proliferator-activated receptor delta (PPARδ) is a nuclearreceptor that is capable of regulating mitochondria biosynthesis. Asshown in PCT/2014/033088, incorporated herein by reference, modulatingthe activity of PPARδ is useful for the treatment of diseases,developmental delays, and symptoms related to mitochondrial dysfunction,such as Alpers Disease, MERRF-Myoclonic epilepsy and ragged-red fiberdisease, Pearson Syndrome, and the like. Modulation PPARδ activity iseffective in the treatment of other conditions, such as musculardiseases, demyelinating diseases, vascular diseases, and metabolicdiseases. Indeed, PPARδ is an important biological target for compoundsused to help treat and prevent mitochondrial diseases, muscle-relateddiseases and disorders, and other related conditions.

Compound A of Formula (I) and Compound B of Formula (II) are PPARδagonists. There is a need for salt forms of these compounds that arecrystalline and otherwise have physical properties that are amenable tolarge scale manufacture. There is also a need for pharmaceuticalformulations in which these drug candidates are stable and areeffectively delivered to the patient.

SUMMARY OF THE INVENTION

Provided herein, inter alia, are salts of Compound A and Compound B andcompositions comprising such compounds that are useful for increasingPPARδ activity.

In one embodiment, provided herein is Compound A of the Formula (I):

in the form of a hemisulfate salt. In one embodiment, the hemisulfatesalt of Compound A is crystalline. Thus, in one embodiment, thecrystalline the hemisulfate salt of Compound A is characterized by anX-ray powder diffraction pattern substantially in accordance with FIG. 1or FIG. 2 .

In another embodiment, provided herein is Compound B of the Formula(II):

in the form of a meglumine salt or a hydrated form of the megluminesalt.

Pharmaceutical compositions of the salts of Compound A and Compound Balso are disclosed herein. Particular embodiments comprise apharmaceutically acceptable carrier or excipient and one or more of thedisclosed compounds. The pharmaceutical compositions of the inventioncan be used in therapy, e.g., for treating a PPARδ-related disease orcondition in a subject.

Another embodiment comprises treating a PPARδ-related disease orcondition in a subject by administering to the subject a therapeuticallyeffective amount of one or both of the disclosed compounds, or apharmaceutical composition comprising the compound(s).

Also provided herein is the use of one or more of the disclosedcompounds, or a pharmaceutical composition comprising one or both of thedisclosed compounds, for the preparation of a medicament for thetreatment of a PPARδ-related disease or condition.

In another embodiment provided herein, the disclosed compounds or apharmaceutical composition comprising one or both of the disclosedcompounds are for use in treating a PPARδ-related disease or condition.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts the X-ray powder diffraction pattern of Compound Ahemisulfate form 1.

FIG. 2 depicts the X-ray powder diffraction pattern of Compound Ahemisulfate form 2.

DETAILED DESCRIPTION OF THE INVENTION

Peroxisome proliferator-activated receptor delta (PPAR-δ), also known asperoxisome proliferator-activated receptor beta (PPAR-β) or as NR1C2(nuclear receptor subfamily 1, group C, member 2), refers to a nuclearreceptor protein that functions as a transcription factor regulating theexpression of genes. PPARS (OMIM 600409) sequences are publicallyavailable, for example from GenBank® sequence database (e.g., accessionnumbers NP_001165289.1 (human, protein) NP_035275 (mouse, protein),NM_001171818 (human, nucleic acid) and NM_011145 (mouse, nucleic acid)).

Ligands of PPARδ, such as Compound A and Compound B, can promotemyoblast proliferation after injury, such as injury to skeletal muscle.As such, as shown in PCT/2014/033088, incorporated herein by reference,modulating the activity of PPARδ is useful for the treatment ofdiseases, developmental delays, and symptoms related to mitochondrialdysfunction, such as Alpers Disease, MERRF-Myoclonic epilepsy andragged-red fiber disease, Pearson Syndrome, and the like. ModulationPPARδ activity is effective in the treatment of other conditions, suchas muscular diseases, demyelinating diseases, vascular diseases, andmetabolic diseases. Indeed, PPARδ is an important biological target forcompounds used to help treat and prevent mitochondrial diseases,muscle-related diseases and disorders, and other related conditions.

Herein, the phrase “PPARδ agonist” refers to substances that increasethe activity of PPARδ. Substances can be tested for their PPARδ agonistactivity by contacting the substance with cells expressing PPARδ,detecting their binding with PPARδ and then detecting signals that serveas the indicator of the activation of PPARδ. Example 1a provides anassay showing that Compound A and Compound B activate PPARδ.

Compounds of the Invention

Provided herein is a hemisulfate salt of(R)-3-methyl-6-(2-((5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-imidazol-1-yl)methyl)phenoxy)hexanoicacid, i.e., Compound A of the Formula (I):

In some embodiments, the hemisulfate salt of Compound A is crystalline.

Also provided herein are methods of making a hemisulfate salt ofCompound A, particularly a crystalline hemisulfate salt of Compound A.For example, upon formation by a reaction between Compound A andsulfuric acid in acetonitrile or 2-propanol, the hemisulfate salt of thecompound can be isolated from the reaction mixture by crystallization(see, e.g., Example 3). Accordingly, in one embodiment, provided hereinis a method of making the hemisulfate salt of Compound A, the methodcomprising the step of reacting Compound A, with sulfuric acid in asolvent to form the hemisulfate salt of Compound A. In a particularembodiment, the solvent comprises acetonitrile. Alternatively, thesolvent comprises 2-propanol. The synthesis of Compound A is describedin Example 2a.

Also provided herein is a meglumine salt of(R)-3-methyl-6-(2-((5-methyl-2-(6-(trifluoromethyl)pyridin-3-yl)-1H-imidazol-1-yl)methyl)phenoxy)hexanoicacid, i.e., Compound B of the Formula (II):

Compound B may also be provided as a hydrate of the meglumine salt. In aparticular embodiment, the meglumine salt of Compound B is provided inmonohydrate form, i.e., the meglumine salt of Compound B is complexedwith water in a one-to-one molar ratio. In other embodiments, themeglumine salt of Compound B is provided in unhydrated form. The term“unhydrated form” means substantially no water is complexed with thecompound, e.g., less than 0.05 equivalents and preferably less than 0.01equivalents of water relative to the compound.

Also provided are methods of making a meglumine salt of Compound B. Forexample, upon formation by a reaction between Compound B and megluminein a solvent such as 2-propanol or acetonitrile, the meglumine salt ofCompound B can be isolated from the reaction mixture (see, e.g., Example11).

Also provided are methods of making a hydrate of the meglumine salt ofCompound B. For example, upon formation by a reaction between Compound Band meglumine in an aqueous solvent mixture such as tetrahydrofuan andwater, the hydrate of the meglumine salt of Compound B can be isolatedfrom the reaction mixture (see, e.g., Example 12).

The synthesis of Compound B is described in Example 2b.

Polymorphic Forms of the Hemisulfate Salt of Compound A

The hemisulfate salt of Compound A can exist in one of at least twopolymorphic forms, i.e., Compound A hemisulfate form 1 and Compound Ahemisulfate form 2. Compound A hemisulfate form 1 possesses acceptablecrystallinity and melting point (Example 6); stability andhygroscopicity (Example 10); and solubility and form control (Example7). As shown in Example 10, Compound A hemisulfate form 1 was determinedto be more thermodynamically stable than Compound A hemisulfate form 2.

Compound A hemisulfate form 1 is characterized by an X-ray powderdiffraction pattern substantially in accordance with FIG. 1 .Specifically, Compound A hemisulfate form 1 is characterized by a X-raypowder diffraction pattern comprising one or more characteristic peaksexpressed in degrees-2-theta (±0.2) as listed in the Table 3 (Example6).

In an embodiment, Compound A hemisulfate form 1 is characterized by afirst X-ray powder diffraction pattern comprising characteristic peaksexpressed in having peaks expressed in degrees-2-theta at angles7.3±0.2°, 14.7±0.2°, 19.1±0.2°, and 22.3±0.2°. In one embodiment, thisfirst X-ray powder diffraction pattern further comprises characteristicpeaks expressed in having peaks expressed in degrees-2-theta one or moreof angles 8.3±0.2°, 15.8±0.2°, 16.5±0.2°, 19.7±0.2°, or 25.8±0.2°. In acertain embodiment, this first X-ray powder diffraction patterncomprising characteristic peaks expressed in having peaks expressed indegrees-2-theta at angles 7.3±0.2°, 8.3±0.2°, 14.7±0.2°, 15.8±0.2°,16.5±0.2°, 19.1±0.2°, 19.7±0.2°, 22.3±0.2°, and 25.8±0.2°. This firstX-ray powder diffraction pattern can also further comprise an X-raypowder diffraction pattern having peaks expressed in degrees-2-theta atone or more of angles 13.0±0.2°, 17.3±0.2°, 23.8±0.2°, 24.5±0.2°,24.9±0.2°, 26.3±0.2°, or 27.8±0.2°. In a certain embodiment, this firstX-ray powder diffraction pattern comprising characteristic peaksexpressed in having peaks expressed in degrees-2-theta at angles7.3±0.2°, 8.3±0.2°, 13.0±0.2°, 14.7±0.2°, 15.8±0.2°, 16.5±0.2°,17.3±0.2°, 19.1±0.2°, 19.7±0.2°, 22.3±0.2°, 23.8±0.2°, 24.5±0.2°,24.9±0.2°, 25.8±0.2°, 26.3±0.2°, and 27.8±0.2°.

In a specific embodiment, this first X-ray powder diffraction patterncomprises peaks expressed in degrees-2-theta at angles 7.3±0.2°,8.3±0.2°, 13.0 0.2°, 14.7±0.2°, 15.8±0.2°, 16.5±0.2°, 17.3±0.2°,19.2±0.2°, 19.7±0.2°, 20.7±0.2°, 22.3±0.2°, 23.8±0.2°, 24.5±0.2°,24.9±0.2°, 25.8±0.2°, 26.3±0.2°, 27.8±0.2°, 28.5±0.2°, 29.6±0.2°, and33.7±0.2°.

Compound A hemisulfate form 2 is characterized by an X-ray powderdiffraction pattern substantially in accordance with FIG. 2 .Specifically, Compound A hemisulfate form 2 is characterized by a X-raypowder diffraction pattern comprising one or more characteristic peaksexpressed in degrees-2-theta (±0.2) as listed in the Table 5 (Example8).

In an embodiment, Compound A hemisulfate form 2 can be characterized bya second X-ray powder diffraction pattern comprising characteristicpeaks expressed in having peaks expressed in degrees-2-theta at angles6.7±0.2°, 13.5±0.2°, 17.4±0.2°, and 18.1±0.2°. In one embodiment, thissecond X-ray powder diffraction pattern further comprises characteristicpeaks expressed in having peaks expressed in degrees-2-theta one or moreof angles 14.5±0.2°, 16.±0.2°, 22.4±0.2°, 23.2, or 23.4±0.2°. In acertain embodiment, this second X-ray powder diffraction patterncomprises characteristic peaks expressed in having peaks expressed indegrees-2-theta at angles 6.7±0.2°, 13.5±0.2°, 14.5±0.2°, 16.±0.2°,17.4±0.2°, 18.1±0.2°, 22.4±0.2°, 23.2, and 23.4±0.2°.

This second X-ray powder diffraction pattern can also further comprisean X-ray powder diffraction pattern having peaks expressed indegrees-2-theta at one or more of angles 10.1±0.2°, 11.1±0.2°,14.2±0.2°, 14.8±0.2°, 16.9±0.2°, 19.0±0.2°, 25.0±0.2°, 26.8±0.2°, or27.4±0.2°. In a certain embodiment, this second X-ray powder diffractionpattern comprises characteristic peaks expressed in having peaksexpressed in degrees-2-theta at angles 6.7±0.2°, 10.1±0.2°, 11.1±0.2°,13.5±0.2°, 14.2±0.2°, 14.5±0.2°, 14.8±0.2°, 16.±0.2°, 16.9±0.2°,17.4±0.2°, 18.1±0.2°, 19.0±0.2°, 22.4±0.2°, 23.2, 23.4±0.2°, 25.0±0.2°,26.8±0.2°, and 27.4±0.2°.

In a specific embodiment, the second X-ray powder diffraction patterncomprises peaks expressed in degrees-2-theta at angles 6.7±0.2°,10.1±0.2°, 11.1±0.2°, 13.5±0.2°, 14.2±0.2°, 14.5±0.2°, 14.8±0.2°,16.1±0.2°, 16.9±0.2°, 17.4±0.2°, 18.1±0.2°, 19.0±0.2°, 19.9±0.2°,22.4±0.2°, 23.2±0.2°, 23.4±0.2°, 25.0±0.2°, 26.8±0.2°, 27.4±0.2°, and29.4±0.2°.

In one embodiment, the crystalline hemisulfate salt of Compound A issubstantially free from impurities. In another embodiment, thecrystalline hemisulfate salt of Compound A comprises less than 10% byweight total impurities. In another embodiment, provided herein is thecrystalline hemisulfate salt of Compound A comprises less than 5% byweight total impurities. In another embodiment, the crystallinehemisulfate salt of Compound A comprises less than 1% by weight totalimpurities. In yet another embodiment, the crystalline hemisulfate saltof Compound A comprises less than 0.1% by weight total impurities.

In certain embodiments, the X-ray powder diffraction pattern of thecrystalline hemisulfate salt of Compound A is collected using Cu K alpha(1.5406 Angstrom) radiation.

In another embodiment, provided herein is the crystalline hemisulfatesalt of Compound A that is substantially free from amorphous hemisulfatesalt of Compound A. As used herein, the term “substantially free fromamorphous hemisulfate salt of Compound A” means that the crystallinehemisulfate salt of Compound A contains no significant amount ofamorphous hemisulfate salt of Compound A. In certain embodiments, atleast about 90% by weight of the crystalline hemisulfate salt ofCompound A is free from amorphous hemisulfate salt of Compound A. Inother embodiments, at least about 95% by weight of the crystallinehemisulfate salt of Compound A is free from amorphous hemisulfate saltof Compound A. In yet other embodiments, at least about 99% by weight ofthe crystalline hemisulfate salt of Compound A is free from amorphoushemisulfate salt of Compound A. In still other embodiments, at leastabout 99.9% by weight of the crystalline hemisulfate salt of Compound Ais free from amorphous hemisulfate salt of Compound A.

In another embodiment, provided herein is the crystalline hemisulfatesalt of Compound A substantially free from other crystalline forms ofhemisulfate salt of Compound A. As used herein, the term “substantiallyfree from other crystalline forms of hemisulfate salt of Compound A”means that the crystalline Compound A contains no significant amount ofother crystalline forms of hemisulfate salt of Compound A. In certainembodiments, at least about 90% by weight of the crystalline hemisulfatesalt of Compound A is free of other crystalline forms.

In other embodiments, at least about 95% by weight of the crystallinehemisulfate salt of Compound A is free of other crystalline forms. Inyet other embodiments, at least about 99% by weight of the crystallinehemisulfate salt of Compound A is free of other crystalline forms. Instill other embodiments, at least about 99.9% by weight of thecrystalline hemisulfate salt of Compound A is free of other crystallineforms.

Methods of Treatment

Methods of treating a PPARδ-related disease or condition in a subjectare disclosed. The methods can include administering to the subject atherapeutically effective amount of one or more compounds orcompositions provided herein.

In one embodiment, the PPARδ-related disease is a mitochondrial disease.Examples of mitochondrial diseases include, but are not limited to,Alpers Disease, CPEO-Chronic progressive external ophthalmoplegia,Kearns-Sayra Syndrome (KSS), Leber Hereditary Optic Neuropathy (LHON),MELAS-Mitochondrial myopathy, encephalomyopathy, lactic acidosis, andstroke-like episodes, MERRF-Myoclonic epilepsy and ragged-red fiberdisease, NARP-neurogenic muscle weakness, ataxia, and retinitispigmentosa, and Pearson Syndrome.

In other embodiments, the PPARδ-related disease is a vascular disease(such as a cardiovascular disease or any disease that would benefit fromincreasing vascularization in tissues exhibiting impaired or inadequateblood flow). In other embodiments, the PPARδ-related disease is amuscular disease, such as a muscular dystrophy. Examples of musculardystrophy include but are not limited to Duchenne muscular dystrophy,Becker muscular dystrophy, limb-girdle muscular dystrophy, congenitalmuscular dystrophy, facioscapulohumeral muscular dystrophy, myotonicmuscular dystrophy, oculopharyngeal muscular dystrophy, distal musculardystrophy, and Emery-Dreifuss muscular dystrophy.

In some embodiments, the PPARδ-related disease or condition is ademyelinating disease, such as multiple sclerosis, Charcot-Marie-Toothdisease, Pelizaeus-Merzbacher disease, encephalomyelitis, neuromyelitisoptica, adrenoleukodystrophy, or Guillian-Barre syndrome.

In other embodiments, the PPARδ-related disease is a metabolic disease.Examples of metabolic diseases include but are not limited to obesity,hypertriglyceridemia, hyperlipidemia, hypoalphalipoproteinemia,hypercholesterolemia, dyslipidemia, Syndrome X, and Type II diabetesmellitus.

In yet other embodiments, the PPARδ-related disease is a musclestructure disorder.

Examples of a muscle structure disorders include, but are not limitedto, Bethlem myopathy, central core disease, congenital fiber typedisproportion, distal muscular dystrophy (MD), Duchenne & Becker MD,Emery-Dreifuss MD, facioscapulohumeral MD, hyaline body myopathy,limb-girdle MD, a muscle sodium channel disorders, myotonicchondrodystrophy, myotonic dystrophy, myotubular myopathy, nemaline bodydisease, oculopharyngeal MD, and stress urinary incontinence.

In still other embodiments, the PPARδ-related disease is a neuronalactivation disorder, Examples of neuronal activation disorders include,but are not limited to, amyotrophic lateral sclerosis,Charcot-Marie-Tooth disease, Guillain-Barre syndrome, Lambert-Eatonsyndrome, multiple sclerosis, myasthenia gravis, nerve lesion,peripheral neuropathy, spinal muscular atrophy, tardy ulnar nerve palsy,and toxic myoneural disorder.

In other embodiments, the PPARδ-related disease is a muscle fatiguedisorder. Examples of muscle fatigue disorders include, but are notlimited to chronic fatigue syndrome, diabetes (type I or II), glycogenstorage disease, fibromyalgia, Friedreich's ataxia, intermittentclaudication, lipid storage myopathy, MELAS, mucopolysaccharidosis,Pompe disease, and thyrotoxic myopathy.

In some embodiments, the PPARδ-related disease is a muscle massdisorder. Examples of muscle mass disorders include, but are not limitedto, cachexia, cartilage degeneration, cerebral palsy, compartmentsyndrome, critical illness myopathy, inclusion body myositis, muscularatrophy (disuse), sarcopenia, steroid myopathy, and systemic lupuserythematosus.

In other embodiments, the PPARδ-related disease is a beta oxidationdisease. Examples of beta oxidation diseases include, but are notlimited to, systemic carnitine transporter, carnitinepalmitoyltransferase (CPT) II deficiency, very long-chain acyl-CoAdehydrogenase (LCHAD or VLCAD) deficiency, trifunctional enzymedeficiency, medium-chain acyl-CoA dehydrogenase (MCAD) deficiency,short-chain acyl-CoA dehydrogenase (SCAD) deficiency, andriboflavin-responsive disorders of β-oxidation (RR-MADD).

In some embodiments, the PPARδ-related disease is a vascular disease.Examples of vascular diseases include, but are not limited to,peripheral vascular insufficiency, peripheral vascular disease,intermittent claudication, peripheral vascular disease (PVD), peripheralartery disease (PAD), peripheral artery occlusive disease (PAOD), andperipheral obliterative arteriopathy.

In other embodiments, the PPARδ-related disease is an ocular vasculardisease. Examples of ocular vascular diseases include, but are notlimited to, age-related macular degeneration (AMD), stargardt disease,hypertensive retinopathy, diabetic retinopathy, retinopathy, maculardegeneration, retinal haemorrhage, and glaucoma.

In yet other embodiments, the PPARδ-related disease is a muscular eyedisease. Examples of muscular eye diseases include, but are not limitedto, strabismus (crossed eye/wandering eye/walleye ophthalmoparesis),progressive external ophthalmoplegia, esotropia, exotropia, a disorderof refraction and accommodation, hypermetropia, myopia, astigmatism,anisometropia, presbyopia, a disorders of accommodation, or internalophthalmoplegia.

In yet other embodiments, the PPARδ-related disease is a metabolicdisease. Examples of metabolic disorders include, but are not limitedto, hyperlipidemia, dyslipidemia, hyperchlolesterolemia,hypertriglyceridemia, HDL hypocholesterolemia, LDL hypercholesterolemiaand/or HLD non-cholesterolemia, VLDL hyperproteinemia,dyslipoproteinemia, apolipoprotein A-I hypoproteinemia, atherosclerosis,disease of arterial sclerosis, disease of cardiovascular systems,cerebrovascular disease, peripheral circulatory disease, metabolicsyndrome, syndrome X, obesity, diabetes (type I or II), hyperglycemia,insulin resistance, impaired glucose tolerance, hyperinsulinism,diabetic complication, cardiac insufficiency, cardiac infarction,cardiomyopathy, hypertension, non-alcoholic fatty liver disease (NAFLD),nonalcoholic steatohepatitis (NASH), thrombus, Alzheimer disease,neurodegenerative disease, demyelinating disease, multiple sclerosis,adrenal leukodystrophy, dermatitis, psoriasis, acne, skin aging,trichosis, inflammation, arthritis, asthma, hypersensitive intestinesyndrome, ulcerative colitis, Crohn's disease, and pancreatitis.

In still other embodiments, the PPARδ-related disease is cancer.Examples of cancer include, but are not limited to, cancers of thecolon, large intestine, skin, breast, prostate, ovary, and/or lung.

In other embodiments, the PPARδ-related disease is a renal disease.Examples of renal diseases include, but are not limited to,glomerulonephritis, glomerulosclerosis, nephrotic syndrome, hypertensivenephrosclerosis, acute nephritis, recurrent hematuria, persistenthematuria, chronic nephritis, rapidly progressive nephritis, acutekidney injury (also known as acute renal failure), chronic renalfailure, diabetic nephropathy, or Bartter's syndrome. PCT/US2014/033088,incorporated herein by reference, demonstrates genetic andpharmacological activation of PPARδ promotes muscle regeneration in anacute thermal injury mouse model. Accordingly, use of PPARδ as atherapeutic target to enhance regenerative efficiency of skeletal muscleis also provided.

Pharmaceutical Compositions and Administration Thereof

The precise amount of compound administered to provide a“therapeutically effective amount” to the subject will depend on themode of administration, the type, and severity of the disease, and onthe characteristics of the subject, such as general health, age, sex,body weight, and tolerance to drugs. The skilled artisan will be able todetermine appropriate dosages depending on these and other factors. Theterm “therapeutically effective amount” means an amount whenadministered to the subject which results in beneficial or desiredresults, including clinical results, e.g., inhibits, suppresses orreduces the symptoms of the condition being treated in the subject ascompared to a control. For example, a therapeutically effective amountcan be from, e.g., 0.1 mg to about 50 g per day.

The terms “administer”, “administering”, “administration”, and the like,as used herein, refer to methods that may be used to enable delivery ofcompositions to the desired site of biological action. These methodsinclude, but are not limited to, intraarticular (in the joints),intravenous, intramuscular, intratumoral, intradermal, intraperitoneal,subcutaneous, orally, topically, intrathecally, inhalationally,transdermally, rectally, and the like. Oral and intravenousadministration are commonly used, for example, when the condition beingtreated is acute kidney injury. Administration techniques that can beemployed with the agents and methods described herein are found in e.g.,Goodman and Gilman, The Pharmacological Basis of Therapeutics, currented.; Pergamon; and Remington's, Pharmaceutical Sciences (currentedition), Mack Publishing Co., Easton, Pa.

Pharmaceutical compositions are disclosed that include the salts ofCompound A and/or Compound B, and typically at least one additionalsubstance, such as an excipient, a known therapeutic other than those ofthe present disclosure, and combinations thereof.

The pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. In an embodiment,the composition is formulated in accordance with routine procedures as apharmaceutical composition adapted for intravenous, subcutaneous,intramuscular, oral, intranasal, or topical administration to humanbeings.

Administration of therapeutic agents by intravenous formulation is wellknown in the pharmaceutical industry. Intravenous formulations comprisethe pharmaceutically active agent dissolved in a pharmaceuticallyacceptable solvent or solution, such as sterile water, normal salinesolutions, lactated Ringer's, or other salt solutions such as Ringer'ssolution.

For example, the formulation should promote the overall stability of theactive ingredient(s), also, the manufacture of the formulation should becost-effective. All of these factors ultimately determine the overallsuccess and usefulness of an intravenous formulation.

An oral formulation typically is prepared as a compressed preparationin, for example, the form of a tablet or pill. A tablet may contain, forexample, about 5-10% of the active ingredient (e.g., a salt of CompoundA or B); about 80% of fillers, disintegrants, lubricants, glidants, andbinders; and 10% of compounds which ensure easy disintegration,disaggregation, and dissolution of the tablet in the stomach or theintestine. Pills can be coated with sugar, varnish, or wax to disguisethe taste.

EXEMPLIFICATION Example 1 PPARδ Activity Screen

Cell Culture and Transfection: CV-1 cells were grown in DMEM+10%charcoal stripped FCS. Cells were seeded into 384-well plates the daybefore transfection to give a confluency of 50-80% at transfection. Atotal of 0.8 g DNA containing 0.64 micrograms pCMX-PPARDelta LBD, 0.1micrograms pCMX.beta.Gal, 0.08 micrograms pGLMH2004 reporter and 0.02micrograms pCMX empty vector was transfected per well using FuGenetransfection reagent according to the manufacturer's instructions(Roche). Cells were allowed to express protein for 48 h followed byaddition of compound.

Plasmids: Human PPARδ was used to PCR amplify the PPARδ LBD. Theamplified cDNA ligand binding domain (LBD) of PPARδ isoform was (PPARδamino acid 128 to C-terminus) and fused to the DNA binding domain (DBD)of the yeast transcription factor GAL4 by subcloning fragments in frameinto the vector pCMX GAL (Sadowski et al. (1992), Gene 118, 137)generating the plasmids pCMX-PPARDelta LBD. Ensuing fusions wereverified by sequencing. The pCMXMH2004 luciferase reporter containsmultiple copies of the GAL4 DNA response element under a minimaleukaryotic promoter (Hollenberg and Evans, 1988). pCMXPGal wasgenerated.

Compounds: All compounds were dissolved in DMSO and diluted 1:1000 uponaddition to the cells. Compounds were tested in quadruple inconcentrations ranging from 0.001 to 100 μM. Cells were treated withcompound for 24 h followed by luciferase assay. Each compound was testedin at least two separate experiments.

Luciferase assay: Medium including test compound was aspirated andwashed with PBS. 50 μl PBS including 1 mM Mg++ and Ca++ were then addedto each well. The luciferase assay was performed using the LucLite kitaccording to the manufacturer's instructions (Packard Instruments).Light emission was quantified by counting on a Perkin Elmer Envisionreader. To measure 3-galactosidase activity 25 μl supernatant from eachtransfection lysate was transferred to a new 384 microplate.Beta-galactosidase assays were performed in the microwell plates using akit from Promega and read in a Perkin Elmer Envision reader. Thebeta-galactosidase data were used to normalize (transfection efficiency,cell growth etc.) the luciferase data.

Statistical Methods: The activity of a compound is calculated as foldinduction compared to an untreated sample. For each compound theefficacy (maximal activity) is given as a relative activity compared toGW501516, a PPARδ agonist. The EC₅₀ is the concentration giving 50% ofmaximal observed activity. EC₅₀ values were calculated via non-linearregression using GraphPad PRISM (GraphPad Software, San Diego, Calif.).

TABLE 1 PPARdelta Activity Screen PPAR delta transactivation EC50Compound Structure Mol. Wt (nM) Compound A

460.41 0.10 Compound B

461.49 4.40

The compounds of this invention show a good agonistic activity of PPARδ,a good selectivity of PPARδ, a good pharmacological effect, good PKprofiles and/or low toxicity including CYP inhibition and hERGinhibition.

Example 2 Synthetic Preparation of Compounds A and B Abbreviations

-   Me methyl-   Et ethyl-   nPr n-propyl-   iPr isopropyl-   cPr cyclopropyl-   nBu n-butyl-   iBu isobutyl-   tBu tert-butyl-   Boc tert-butyloxycarbonyl-   Ac acetyl-   Ph phenyl-   Tf trifluoromethanesulfonyl-   Ts 4-methylphenylsulfonyl-   DIAD diisopropyl azodicarboxylate-   EDCI 3-(3-dimethylaminopropyl)-1-ethylcarbodiimide-   HOBt 1-hydroxybenzotriazole-   HATU    1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium    3-oxide hexafluorophosphate-   HBTU N,N,N,N′,N′-Tetramethyl-O-(1H-benzotriazol-1-yl)uronium    hexafluorophosphate-   NBS N-bromosuccinimide-   DIPEA diisopropylethylamine-   mCPBA m-chloroperoxybenzoic acid-   Togni's reagent 3,3-dimethyl-1-(trifluoromethyl)-1,2-benziodoxole-   DCM dichloromethane-   DME dimethoxyethane-   DMF N,N-dimethylformamide-   DMF·DMA N,N-dimethylformamide dimethyl acetal-   DMSO dimethylsulfoxide-   TFA trifluoroacetic acid-   THF tetrahydrofuran-   MW microwave irradiation-   aq Aqueous-   M concentration expressed in mol/L-   RT room temperature-   TLC thin layer chromatography-   HPLC high-performance liquid chromatography-   MPLC medium pressure liquid chromatography-   LCMS liquid chromatography-mass spectrometry-   ESI+ Electrospray ionization positive mode-   ESI− Electrospray ionization negative mode-   ¹H NMR (DMSO-d₆) δ (ppm) of peak in H NMR in DMSO-d₆-   s singlet (spectrum)-   d doublet (spectrum)-   t triplet (spectrum)-   q quartet (spectrum)-   dd double doublet (spectrum)-   br broad line (spectrum)-   m multiplet (spectrum)

Example 2a: Synthesis of(R)-3-methyl-6-(2-((5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-imidazol-1-yl)methyl)phenoxy)hexanoicacid (Compound A)

Synthesis of ethyl (R)-6-bromo-3-methylhexanoate

In a 1 L round bottom flask, a solution of ethyl(R)-6-hydroxy-3-methylhexanoate (65.0 g, 373.56 mmol) in DCM (650 mL)was treated with PBr₃ (101.0 g, 373.56 mmol) at RT. The reaction mixturewas stirred at RT for 3 h. Upon completion of reaction (monitored byTLC), the reaction mixture was diluted with water (500 mL) and extractedwith diethyl ether (3×500 mL). The organic extract was separated anddried over anhydrous Na₂SO₄. The solvent was removed under reducedpressure to get the title compound (57.12 g).

Step-1: Synthesis of N-(prop-2-yn-1-yl)-4-(trifluoromethyl)benzamide

In a 500 mL round bottom flask, a stirred solution of4-(trifluoromethyl)benzoic acid (10 g, 52.63 mmol) and prop-2-yn-1-amine(3.47 g, 63.15 mmol) in DMF (200 mL) was treated sequentially withEDCI·HCl (20.09 g, 105.2 mmol), HOBt (14.2 g, 105.2 mmol) and Et₃N (14.6mL, 105.2 mmol) at RT under nitrogen atmosphere. The reaction mixturewas stirred at RT for 12 h under nitrogen atmosphere. Upon completion ofreaction (monitored by TLC), the reaction mixture was diluted with icecold water and solid precipitated out. The solid was filtered and driedunder reduced pressure to yield the title compound (8.42 g, 70.5%).

¹H NMR (300 MHz, CDCl₃): δ 7.90 (d, J=8.1 Hz, 2H), 7.71 (d, J=8.8 Hz,2H), 6.47 (brs, 1H), 4.28-4.62 (m, 2H), 3.12 (t, J=2.4 Hz, 1H). LCMS(ESI+, m/z): 228.2 (M+H)⁺.

Step-2: Synthesis of1-(2-methoxybenzyl)-5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-imidazole

In a 500 mL resealable reaction tube, a solution ofN-(prop-2-yn-1-yl)-4-(trifluoromethyl)benzamide (13.3 g, 58.59 mmol) and2-methoxybenzyl amine (12.0 g, 87.84 mmol) in toluene (150 mL) wastreated with Zn(OTf)₂ (6.67 g, 17.5 mmol) at RT under nitrogenatmosphere. The reaction mixture was heated at 110° C. for 12 h. Uponcompletion of reaction (monitored by TLC), the reaction mixture wasdiluted with water and extracted with EtOAc (3×100 mL). The combinedorganic extract was washed with saturated NaHCO₃, brine and dried overanhydrous Na₂SO₄. The solution was concentrated under reduced pressureand the residue obtained was purified by silica gel columnchromatography (elution, 25% EtOAc in hexanes) to afford the titlecompound (17.3 g, 85.3%).

¹H NMR (400 MHz, CDCl₃): δ 7.59-7.54 (m, 4H), 7.30-7.23 (m, 1H), 7.00(s, 1H), 6.91-6.86 (m, 2H), 6.57 (d, J=7.2 Hz, 1H), 5.11 (s, 2H), 3.84(s, 3H), 2.11 (s, 3H). LCMS (ESI+, m/z): 347.3 (M+H)⁺.

Step-3: Synthesis of2-((5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-imidazol-1-yl)methyl)phenol

In a 500 mL round bottom flask, a solution of1-(2-methoxybenzyl)-5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-imidazole(17.3 g, 49.94 mmol) in DCM (150 mL) was treated with BBr₃ (1.0 M, 90.0mL) drop wise at 0° C. The reaction mixture was stirred at RT for 4 h.Upon completion of reaction (monitored by TLC), the reaction mixture wasbasified (pH˜9) with aqueous NaHCO₃ and extracted with EtOAc (3×500 mL).The combined organic extract was dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to afford the title compound (19.2g, crude).

¹H NMR (400 MHz, DMSO-d₆): δ 9.99 (s, 1H), 7.88 (d, J=8.4 Hz, 2H), 7.77(d, J=8.4 Hz, 2H), 7.33 (s, 1H), 7.14-7.10 (m, 1H), 6.83 (d, J=8.0 Hz,1H), 6.74-6.70 (m, 1H), 6.55 (d, J=6.8 Hz, 1H), 5.21 (s, 2H), 2.16 (s,3H). LCMS (ESI+, m/z): 333.3 (M+H)⁺.

Step-4: Synthesis of ethyl(R)-3-methyl-6-(2-((5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-imidazol-1-yl)methyl)phenoxy)hexanoate

In a 250 mL round bottom flask, a stirred solution of2-((5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-imidazol-1-yl)methyl)phenol (4.0 g, 12.0 mmol) in DMF (100mL) was treated with KO^(t)Bu (4.03 g, 36.1 mmol) and ethyl(R)-6-bromo-3-methylhexanoate (8.52 g, 36.10 mmol) at RT under nitrogenatmosphere. The resulting reaction mixture was stirred at RT for 12 h.Upon completion of the reaction (monitored by TLC), the reaction mixturequenched with ice cold water and extracted with EtOAc (3×100 mL). Thecombined organic extract was washed with brine, dried over anhydrousNa₂SO₄ and concentrated under reduced pressure. The residue obtained waspurified by silica gel column chromatography (gradient elution, 15-30%EtOAc in hexanes) to afford the title compound (3.31 g, 56.3%). LCMS(ESI+, m/z): 489.3 (M+H)⁺.

Step-5: Synthesis of(R)-3-methyl-6-(2-((5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-imidazol-1-yl)methyl)phenoxy)hexanoicacid (Compound A)

In a 250 mL round bottom flask, a stirred solution of ethyl(R)-3-methyl-6-(2-((5-methyl-2-(4-(trifluoromethyl)phenyl)-1H-imidazol-1-yl)methyl)phenoxy)hexanoate(3.3 g, 6.75 mmol) in THF (30 mL), ethanol (10 mL) and water (10 mL) wastreated with lithium hydroxide monohydrate (1.42 g, 33.8 mmol) at RT.The reaction mixture was stirred at RT for 12 h. Upon completion ofreaction (monitored by TLC), the reaction mixture was concentrated underreduced pressure. The residue obtained was washed with EtOAc, dilutedwith cold water and acidified (pH˜5) with 1N HCl. The solid obtained wasfiltered and dried under reduced pressure to give the title compound(1.12 g, 36.0%).

¹H NMR (400 MHz, DMSO-d₆): δ 12.00 (brs, 1H), 7.71 (d, J=8.4 Hz, 2H),7.62 (d, J=8.4 Hz, 2H), 7.26-7.21 (m, 1H), 7.01 (d, J=8.4 Hz, 1H), 6.93(s, 1H), 6.86-6.83 (m, 1H), 6.38 (d, J=6.8 Hz, 1H), 5.16 (s, 2H), 3.98(t, J=6.0 Hz, 2H), 2.19-2.14 (m, 1H), 2.08 (s, 3H), 1.99-1.93 (m, 1H),1.84-1.76 (m, 1H), 1.67-1.65 (m, 2H), 1.45-1.42 (m, 1H), 1.28-1.18 (m,1H), 0.83 (d, J=6.4 Hz, 3H). ¹⁹F NMR (400 MHz, DMSO-d₆): δ −56.4. LCMS(ESI+, m/z): 460.8 (M+H)⁺. HPLC: 98.89% (210 nm).

Example 2b: Synthesis of(R)-3-methyl-6-(2-((5-methyl-2-(6-(trifluoromethyl)pyridin-3-yl)-1H-imidazol-1-yl)methyl)phenoxy)hexanoicacid (Compound B)

Step-1: Synthesis of N-(prop-2-yn-1-yl)-6-(trifluoromethyl)nicotinamide

In a 100 mL round bottom flask, a stirred solution of6-(trifluoromethyl)nicotinic acid (3 g, 15.70 mmol) andprop-2-yn-1-amine (1.05 g, 18.84 mmol) in DMF (50 mL) was treated withHATU (7.2 g, 18.84 mmol) and Et₃N (3.1 mL, 23.55 mmol) at RT undernitrogen atmosphere. The resulting reaction mixture was stirred at RTfor 3 h. Upon completion of reaction (monitored by TLC), the reactionmixture was diluted with cold water and solid precipitated was filtered,washed with water and dried under reduced pressure to get the titlecompound (2.6 g, 72.6%).

¹H NMR (300 MHz, CDCl₃): δ 9.08 (d, J=2.1 Hz, 1H), 8.32 (dd, J=8.4, 2.4Hz, 1H), 7.78 (d, J=7.8 Hz, 1H), 6.62 (brs, 1H), 4.30-4.28 (m, 2H), 2.33(t, J=2.4 Hz, 1H).

LCMS (ESI+, m/z): 229.2 (M+H)⁺.

Step-2: Synthesis of5-(1-(2-methoxybenzyl)-5-methyl-1H-imidazol-2-yl)-2-(trifluoromethyl)pyridine

In a 50 mL resealable tube, a solution ofN-(prop-2-yn-1-yl)-6-(trifluoromethyl) nicotinamide (1.0 g, 4.38 mmol)and 2-methoxyphenybenzyl amine (1.2 g, 8.77 mmol) in toluene (10 mL) wastreated with Zn(OTf)₂ (0.16 g, 0.43 mmol) at RT under nitrogenatmosphere. The reaction mixture was heated at 120° C. for 12 h. Uponcompletion of reaction (monitored by TLC), the reaction mixture wasdiluted with water and extracted with EtOAc (3×20 mL). The organicextract was washed with saturated NaHCO₃, brine and dried over anhydrousNa₂SO₄. The solution was concentrated under reduced pressure and residueobtained was purified by silica gel column chromatography (elution, 25%EtOAc in hexanes) to yield the title compound (0.8 g, 52.6%).

¹H NMR (400 MHz, CDCl₃): δ 8.79 (s, 1H), 8.07 (d, J=8.1 Hz, 1H), 7.68(d, J=8.1 Hz, 1H), 7.31 (t, J=8.4 Hz, 1H), 7.09 (s, 1H), 6.94-6.87 (m,2H), 6.56 (d, J=7.5 Hz, 1H), 5.16 (s, 2H), 3.87 (s, 3H). LCMS (ESI+,m/z): 348.3 (M+H)⁺.

Step-3: Synthesis of2-((5-methyl-2-(6-(trifluoromethyl)pyridin-3-yl)-1H-imidazol-1-yl)methyl)phenol

In a 100 mL round bottom flask, a solution of5-(1-(2-methoxybenzyl)-5-methyl-1H-imidazol-2-yl)-2-(trifluoromethyl)pyridine(0.8 g, 2.31 mmol) in dichloromethane (300 mL) was treated with BBr₃(0.8 mL, 2.31 mmol) drop wise at 0° C. The reaction mixture was stirredat RT for 2 h. Upon completion of reaction (monitored by TLC), thereaction mixture was basified (pH˜9) with aqueous NaHCO₃ and extractedwith EtOAc. The organic extract was dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to afford the title compound (0.5 g,65.1%)

¹H NMR (400 MHz, DMSO-d₆): δ 9.92 (s, 1H), 8.83 (s, 1H), 8.12 (d, J=8.1Hz, 1H), 7.94 (d, J=8.1 Hz, 1H), 7.12 (d, J=6.9 Hz, 1H), 7.02 (s, 1H),6.87 (d, J=7.8 Hz 1H), 6.73 (t, J=7.2 Hz, 1H), 6.37 (d, J=7.2 Hz, 1H),5.20 (s, 2H), 2.15 (s, 3H). LCMS (ESI+, m/z): 334.3 (M+H)⁺.

Step-4: Synthesis of ethyl(R)-3-methyl-6-(2-((5-methyl-2-(6-(trifluoromethyl)pyridin-3-yl)-1H-imidazol-1-yl)methyl)phenoxy)hexanoate

In a 50 mL round bottom flask, a stirred solution of2-((5-methyl-2-(6-(trifluoromethyl)pyridin-3-yl)-1H-imidazol-1-yl)methyl)phenol(0.5 g, 1.50 mmol) (a procedure for the preparation of which isdisclosed in U.S. Application No. 62/061,483, incorporated herein byreference) in DMF (10 mL) was treated with K₂CO₃ (0.41 g, 3.00 mmol) andethyl (R)-6-bromo-3-methylhexanoate (0.710 g, 3.00 mmol) at RT undernitrogen atmosphere. The resulting reaction mixture was heated 60° C.for 12 h. Upon completion of the reaction (monitored by TLC), thereaction mixture was quenched with ice cold water and extracted withethyl acetate (75 mL×3). The combined organic extract was washed withbrine, dried over anhydrous Na₂SO₄ and concentrated under reducedpressure. The residue obtained was purified by silica gel columnchromatography (gradient elution, 15-30% EtOAc in hexanes) to afford thetitle compound (0.45 g, 61.3%).

LCMS (ESI+, m/z): 491.0 (M+H)⁺.

Step-5: Synthesis of(R)-3-methyl-6-(2-((5-methyl-2-(6-(trifluoromethyl)pyridin-3-yl)-1H-imidazol-1-yl)methyl)phenoxy)hexanoicacid (Compound B)

In a 250 mL round bottom flask, a stirred solution of ethyl(R)-3-methyl-6-(2-((5-methyl-2-(6-(trifluoromethyl)pyridin-3-yl)-1H-imidazol-1-yl)methyl)phenoxy)hexanoate(0.45 g, 0.92 mmol) in THF (5 mL), ethanol (2.5 mL) and water (2.5 mL)was treated with lithium hydroxide monohydrate (16.0 g, 74.33 mmol) atRT. The reaction mixture was stirred at RT for 12 h. Upon completion ofreaction (monitored by TLC), the reaction mixture was concentrated underreduced pressure. The residue obtained was washed with EtOAc, dilutedwith cold water and acidified (pH˜5) with 1N HCl. The solid was filteredand dried under reduced pressure to give the title compound (0.166 g,39.2%).

¹H NMR (400 MHz, DMSO-d₆): δ11.96 (brs, 1H), 8.79 (s, 1H), 8.05 (d,J=8.0 Hz, 1H), 7.90 (d, J=8.0 Hz, 1H), 7.24 (t, J=7.6 Hz, 1H), 7.02 (d,J=8.4 Hz, 1H), 7.00 (s, 1H), 6.84 (t, J=7.6 Hz, 1H), 6.43 (d, J=7.2 Hz,1H), 5.21 (s, 2H), 3.98 (t, J=6.0 Hz, 2H), 2.19-2.14 (m, 1H), 2.13 (s,3H), 2.03-1.94 (m, 1H), 1.85-1.80 (m, 1H), 1.68-1.66 (m, 2H), 1.38-1.36(m, 1H), 1.28-1.18 (m, 1H), 0.85 (d, J=6.4 Hz, 3H). ¹⁹F NMR (400 MHz,DMSO-d₆): δ −66.46.

LCMS (ESI+, m/z): 462.3 (M+H)⁺. HPLC: 95.11% (210 nm).

Example 3 Compound A Salt Screen

A 25 mg/mL stock solution of Compound A was prepared in methanol. Then,2 mL of Compound A stock solution was dispensed into 4 mL amber glassvials. Salt formers (1 eq) were then added to the vials as listed inTable 1 and the solvents were removed by evaporation under a stream ofnitrogen. Once evaporation was completed, the screening solvents listedin Table 1 were pipetted in, the vials were sealed, and the samples wereput onto a 50° C. stir plate to stir for up to an hour. Then, the heatwas shut off and samples were allowed to cool to 25° C. with stirring.Experiments resulting in slurries were stirred. Experiments resulting insolutions were converted to evaporative crystallizations.

Solids isolated from the experiments were characterized by XRPD todetermine if they were crystalline as well as unique solid state formssuggesting salt formation occurred.

An additional experiment was performed with the suspected sodium salt inan attempt to create a Compound A sodium salt. The suspected sodium saltisolated from acetonitrile was slurried in ethyl acetate for six days at25° C. Solids were analyzed by XRPD, which showed a physical mixture ofCompound A free form and the starting suspected sodium salt isolatedfrom acetonitrile. No new salt form was generated.

TABLE 1 Salt Screening Experiments for Compound A Sample Salt FormerSolvent Temperature Results 1 HCl 2-Propanol 25° C. Gel 2 H2SO42-Propanol 25° C. New Salt Form A (Hemisulfate Form 1) 3 H3PO42-Propanol 25° C. New Salt Form B 4 p-Toluenesulfonic Acid 2-Propanol25° C. Gel 5 Methanesulfonic Acid 2-Propanol 25° C. Gel 6 NaOH2-Propanol 25° C. New Salt Form C 7 KOH 2-Propanol 25° C. Gel 8 CholineHydroxide 2-Propanol 25° C. New Salt Form D 9 L-Arginine 2-Propanol 25°C. Amorphous 10 L-Lysine 2-Propanol 25° C. New Salt Form E 11N-Methyl-D-Glucamine 2-Propanol 25° C. Gel 12 TRIS 2-Propanol 25° C. NewSalt Form F 13 HCl Acetonitrile 25° C. Gel 14 H2SO4 Acetonitrile  4° C.New Salt Form A (Hemisulfate Form 1) 15 H3PO4 Acetonitrile 25° C. NewSalt Form B 16 p-Toluenesulfonic Acid Acetonitrile 25° C. Gel 17Methanesulfonic Acid Acetonitrile 25° C. Gel 18 NaOH Acetonitrile 25° C.New Salt Form C 19 KOH Acetonitrile 25° C. Semi-Crystalline 20 CholineHydroxide Acetonitrile 25° C. New Salt Form D 21 L-Arginine Acetonitrile25° C. Amorphous 22 L-Lysine Acetonitrile 25° C. New Salt Form E 23N-Methyl-D-Glucamine Acetonitrile 25° C. Mixture of Starting Material 24TRIS Acetonitrile  4° C. Starting Material

Example 4 Stability of Compound A Salts

The stability of the sulfate, phosphate, L-lysine, and TRIS saltsprepared in Example 3 were tested. Approximately 20 mg of each samplewas combined with 1 mL of water in microcentrifuge tubes. The sampleswere allowed to mix overnight on a temperature controlled shaker at 20°C. at 850 rpm. The solids were characterized by XPRD to determine if asolid state form change occurred. There was no change in the solids forthe suspected sulfate salt as well as the starting material. However,the suspected phosphate, L-lysine, and TRIS salts appear todisproportionate back to the starting material in aqueous environments.

Example 5 Preparation of Compound A Hemisulfate Form 1

In a 50 mL vial was dissolved 883.2 mg of Compound A was dissolved in 35mL methanol. Then, H₂SO₄ (1920 μL, 1M in H₂O, 1 equivalent) was pipettedin. The solvent was allowed to evaporate under N₂. Once evaporated,2-propanol (18 mL) was pipetted in followed by a stir bar. The vial wascapped and placed on a 50° C. stir plate for 1 hour, then thetemperature was dropped to 25° C., where it stirred for 1 day. After 1day, the solids were filtered under vacuum and allowed to air dry.

¹H NMR (400 MHz, DMSO-d₆): δ 7.85 (d, J=8.4 Hz, 2H), 7.74 (d, J=8.4 Hz,2H), 7.36 (s, 1H), 7.27 (t, J=8.4 Hz, 1H), 7.02 (d, J=8.4 Hz, 1H), 6.85(t, J=7.6 Hz, 1H), 6.62 (d, J=7.2 Hz, 1H), 5.26 (s, 2H), 3.96 (t, J=6.0Hz, 2H), 2.21-2.16 (m, 4H), 1.96 (dd, J=8.0, 15.2 Hz, 1H), 1.83-1.80 (m,1H), 1.67-1.59 (m, 2H), 1.35-1.31 (m, 1H), 1.28-1.18 (m, 1H), 0.85 (d,J=6.4 Hz, 3H).

Mass Spectrum (ESI) m/e 461.2.

Elemental Analysis: Calculated: C, 58.93%; H, 5.54%; N, 5.50%; S, 3.15.Observed: C, 58.30%; H, 5.36%; N, 5.42%; S, 3.47.

The Compound A hemisulfate form 1 was also obtained according to thesame manner as that mentioned above using acetonitrile (18 mL) as asolvent instead of 2-propanol (18 mL).

The elemental analysis of Compound A hemisulfate form 1 shows that itcontains a two to one ratio of Compound A cation to sulfate anion.

The presence of the sulfate anion as well as the sulfate stoichiometrywas confirmed through elemental analysis, which revealed two moleculesof Compound A to one molecule of sulfate anion.

The sulfate salt was named Compound A hemisulfate form 1. This form wassubjected to a thermodynamic stability form screen to see if a morestable form could be identified as well as screen for solvates,hydrates, and assess the risk of disproportionation (Example 7).Chemical and physical stability data as well as hygroscopicity data werealso generated for Compound A hemisulfate form 1 (Example 11).

Example 6 Compound A Hemisulfate Form 1 Characterization

Physical characterization consisting of XRPD (FIG. 1 ), TGA, and DSC wasperformed for Compound A hemisulfate form 1. A summary of the XRPD datafrom FIG. 1 for Compound A hemisulfate Form 1 is provided in Table 3.

TABLE 3 Number Position [°2θ] d-spacing [Å] Height [cts] 1 7.272312.15596 203.49 2 8.3321 10.61206 87.18 3 12.9823 6.81943 34.79 4 14.6826.03356 187.57 5 15.7717 5.61908 68.85 6 16.5492 5.35679 72.4 7 17.30665.12403 36.33 8 19.1525 4.63415 149.22 9 19.6624 4.51512 80.9 10 20.71874.28724 15.2 11 22.268 3.99234 224.73 12 23.8209 3.73547 49.9 13 24.53623.62817 37.27 14 24.9107 3.57448 34.45 15 25.7712 3.45704 95.31 1626.2777 3.39155 39.15 17 27.8254 3.20632 40.71 18 28.5033 3.13158 16 1929.5617 3.02183 16.5 20 33.7206 2.65804 3.18

X-ray powder diffraction data were collected under ambient conditions ona Rigaku Miniflex 600 diffractometer using Cu K alpha (1.5406 Angstrom)radiation. Powder patterns were collected on a zero background holderwith a 0.1 mm indent at a scan rate of 2 to 400 two theta at 20 per minat 40 kV and 15 mA. Data was analyzed using High Score Plus version 4.1.

Example 7 Compound A Hemisulfate Form Screening

A thermodynamic stability form screen was initiated with Compound Ahemisulfate form 1 to screen for thermodynamically more stablepolymorphs, solvates, and hydrates as well as to probe the tendency todisproportionate back to Compound A. Approximately 90 to 110 mg ofCompound A hemisulfate form 1 was weighed out and transferred into 4 mLamber glass vials followed by 0.8 to 1 mL of solvent and a magnetic stirbar. The vials were sealed then placed onto temperature controlled stirplates and stirred for fifteen days at 500 rpm. Solvents andtemperatures are listed in Table 4. XRPD analyses of the solids revealedno change in solid state form occurred except for the solids isolatedfrom experiments in methanol at 25° C. and water/methanol at 25° C. Thisnew form is different than the starting Compound A free form andCompound A hemisulfate form 1.

TABLE 4 Compound A Hemisulfate Slurry Screen Experiment SolventTemperature Results 1 Ethyl Acetate 50° C. No change in form 22-Propanol 50° C. No change in form 3 Acetone 25° C. No change in form 4Dichloromethane  4° C. No change in form 5 Acetonitrile 50° C. No changein form 6 Tetrahydrofuran 25° C. No change in form 7 Water 25° C. Nochange in form 8 Water/Methanol 25° C. Compound A (1/3, v/v) HemisulfateForm 2 9 Water/Acetonitrile  4° C. No change in form (1/3, v/v) 10Ethanol 50° C. No change in form 11 2-Methyl-THF 25° C. No change inform 12 CPME 50° C. No change in form 13 Toluene 50° C. No change inform 14 Methanol 25° C. Compound A Hemisulfate Form 2

Example 8 Compound A Hemisulfate Form 2 Characterization

The new solid state form isolated from the methanol slurry andwater/methanol slurry in Example 7 was subjected to additionalcharacterization and found to be a new polymorph of the hemisulfate saltof Compound A. This new form was named Compound A hemisulfate form 2.Physical characterization consisting of XRPD (FIG. 2 ), TGA and DSC wasperformed for Compound A hemisulfate form 2 after drying under nitrogen.A summary of the XRPD data from FIG. 2 for Compound A hemisulfate form 2is provided in Table 5.

TABLE 5 Number Position [°2θ] d-spacing [Å] Height [cts] 1 6.669913.25246 565.65 2 10.0501 8.80157 17.65 3 11.077 7.98776 17.71 4 13.50166.55831 54.27 5 14.2386 6.22046 23.63 6 14.5068 6.10605 44.53 7 14.84455.96788 22.89 8 16.0872 5.50958 33.53 9 16.9192 5.24047 17.12 10 17.43395.08691 84.25 11 18.0914 4.90349 288.69 12 18.975 4.67709 26.04 1319.899 4.46195 12.96 14 22.3828 3.97212 48.96 15 23.2046 3.83327 41.8216 23.4233 3.79797 44.01 17 25.0416 3.55607 29.72 18 26.7503 3.3326920.02 19 27.3864 3.2567 20.05 20 29.3568 3.04246 9.17

X-ray powder diffraction data were collected under ambient conditions ona Rigaku Miniflex 600 diffractometer using Cu K alpha (1.5406 Angstrom)radiation. Powder patterns were collected on a zero background holderwith a 0.1 mm indent at a scan rate of 2 to 400 two theta at 2° per minat 40 kV and 15 mA. Data was analyzed using High Score Plus version 4.1.

Solution ¹HNMR and elemental analysis data were also generated to checkfor chemical changes, as compared against the free base and initialhemisulfate form 1. The data indicate no molecular transformationoccurred and that Compound A hemisulfate form 2 is not a solvate. Also,the solution ¹H NMR and elemental analysis data were consistent with thematerial being a hemisulfate salt having a two to one ration of CompoundA cation to sulfate anion.

Example 9 Relative Thermodynamic Stability of Compound A HemisulfateForm 1 and Form 2

The relative thermodynamic stability of Compound A hemisulfate form 1and hemisulfate form 2 was determined through a comparison of thermalanalysis data for each form as well as competition slurry experiments.DSC data for Compound A hemisulfate form 1 revealed this form has anonset melting point of 185.7° C. and an enthalpy of fusion of 106.9joules per gram. DSC data for Compound A hemisulfate form 2 revealedthis form has an onset melting point of 177.4° C. and an enthalpy offusion of 98.9 joules per gram. According to the heat of fusion rule,Compound A hemisulfate form 1 and hemisulfate form 2 are montropicallyrelated given Compound A hemisulfate form 1 has the higher melting pointand the higher enthalpy of fusion of the two polymorphs. Compound Ahemisulfate form 1 is the thermodynamically more stable form.

Competition slurry experiments were conducted to confirm the relativethermodynamic stabilities of Compound A hemisulfate form 1 and CompoundA hemisulfate form 2. Mixtures of both forms were slurried together in0.5 mL acetone at 25° C. for one week. Both forms were still present inthe slurry, so the experiment was moved to 4° C. After ten days at 4°C., XRPD analysis showed that the mixture converted to Compound Ahemisulfate form 1, indicating Compound A hemisulfate form 1 is morestable than Compound A hemisulfate form 2 at 4° C.

Compound A hemisulfate form 1 and Compound A hemisulfate form 2 wereslurried together in 0.5 mL acetonitrile at 50° C. After three days at50° C., XRPD analysis showed both forms still remaining in the slurry.After ten days at 50° C. in acetonitrile, XRPD analysis showed that themixture converted to Compound A hemisulfate form 1, indicating theCompound A hemisulfate form 1 is more thermodynamically stable thanCompound A hemisulfate form 2 at 50° C.

Compound A hemisulfate form 1 and Compound A hemisulfate form 2 wereslurried together in 0.5 mL ethyl acetate at 25° C. After two days, bothforms were still present in the slurry, so the experiment was moved to4° C. After eight days at 4° C., XRPD analysis showed that the solidsremained a mixture of forms. This experiment was stopped before fullyconverting to a single form given mixtures converted to Compound Ahemisulfate form 1 in acetone at 4° C. and in acetonitrile at 50° C.This experiment demonstrates the conversion from Compound A hemisulfateform 2 to Compound A hemisulfate form 1 can be slow under certainconditions.

Both thermal analysis data and competition slurry experiment datademonstrate Compound A hemisulfate form 1 is thermodynamically morestable than Compound A hemisulfate form 2. Methanol was always presentin the solvent system when Compound A hemisulfate form 2 was formedwhich suggests Compound A hemisulfate form 2 might be formed through thedesolvation of an unstable and never observed methanol solvate.

Example 10 Compound A Hemisulfate Form 1 Stability and Hygroscopicity

An eight week chemical and physical stability study was conducted forCompound A hemisulfate form 1. The material was stored at 4° C., 25°C./60% RH, 40° C., and 40° C./75% RH for eight weeks as well as stressedconditions of 70° C. and 70° C./75% RH for two weeks. In addition,photostability was also measured after exposing the material to twocycles of ICH conditions. Degradation and recovery was measured by HPLC.See Table 6 for HPLC results, including percent recovery of solids andrelative retention time area percentages, compared against the standardstored at 4° C.

TABLE 6 Summary of HPLC data. Sample % RRT *RRT RRT RRT RRT RRT RRT 4°C., 4 wk 101 1.16 0.21 97.36 0.21 0.20 0.25 0.58 4° C., 8 wk 100 1.150.12 97.42 0.34 0.19 0.24 0.54 25° C./60% 100 1.20 0.21 97.40 0.21 0.210.25 0.50 RH, 4 wk 25° C./60% 99 1.15 0.04 97.67 0.25 0.19 0.23 0.46 RH,8 wk 40° C., 4 wk 101 1.20 0.22 97.27 0.21 0.20 0.25 0.59 40° C., 8 wk101 1.16 0.11 97.44 0.33 0.19 0.24 0.53 40° C./75% 101 1.24 0.21 97.330.21 0.21 0.25 0.50 RH, 4 wk 40° C./75% 100 1.19 0.04 97.63 0.30 0.190.23 0.42 RH, 8 wk 70° C., 2 wk 101 1.20 0.18 97.34 0.22 0.21 0.26 0.5570° C./75% 101 1.22 0.19 97.45 0.22 0.21 0.24 0.42 RH, 2 wk Photo 981.11 0.10 97.78 0.27 0.20 0.14 0.40 Control (dark) 100 1.16 0.07 97.530.30 0.19 0.23 0.53 Standard N/A 1.18 0.14 97.38 0.37 0.18 0.23 0.53

XRPD was used to check the physical stability. No changes in solid stateform were observed in all conditions.

Dynamic vapor sorption analysis was performed on Compound A hemisulfateform 1 at 25° C. At approximately 90% RH, the material reversibly picksup approximately 3.5% water by weight. After DVS was completed, thesolid collected was checked by XRPD, which showed that the materialremained Compound A hemisulfate form 1.

Example 11 Preparation of Meglumine Salt of Compound B

Two separate methods were used to generate a meglumine salt of CompoundB.

Method 1

Compound B (102.7 mg) was combined with meglumine (43.7 mg) and 2 mL of2-propanol in a 4 mL glass vial. The vial was sealed with a cap and thecontents were subjected to sonication at 25° C. for 20 minutes followedby stirring at 50° C. for 60 minutes. The vial was then moved to a newstir plate and the slurry in the vial was stirred at 25° C.

Method 2

Compound B (102.2 mg) was combined with meglumine (43.2 mg) and 2 mL ofacetonitrile in a 4 mL glass vial. The vial was sealed with a cap andthe contents were subjected to sonication at 25° C. for 20 minutesfollowed by stirring at 50° C. for 60 minutes. The vial was then movedto a new stir plate and the slurry in the vial was stirred at 25° C.

For both method 1 and method 2, after 2 days of stirring at 25° C., bothsamples were centrifuged, supernatants discarded, and solids were airdried.

Example 12 Preparation of Hydrate of Meglumine Salt of Compound B

In a 500 mL round bottom flask, a stirred solution of Compound B (20 g,43.33 mmol) in THF (100 mL) and water (100 mL) was treated meglumine(8.45 g, 43.33 mmol) at 0° C. The resulting reaction mixture was stirredat RT for 6 h. The reaction mixture was concentrated under reducedpressure and solid obtained was dried under reduced pressure (3 h) toafford the title compound as a white solid (28.5 g, 98.95%).

¹H NMR (400 MHz, CD₃OD): δ 8.75 (s, 1H), 8.02 (d, J=8.4 Hz, 1H), 7.82(d, J=8.0 Hz 1H), 7.26 (t, J=8.4 Hz, 1H), 7.03 (s, 1H), 6.99 (d, J=8 Hz,1H), 6.85 (t, J=7.6 Hz, 1H), 6.50 (d, J=7.6 Hz, 1H), 5.25 (s, 2H),4.09-3.99 (m, 3H), 3.97-3.77 (m, 2H), 3.74-3.61 (m, 3H), 3.29-3.06 (m,2H), 2.64 (s, 3H), 2.22 (s, 3H), 2.18-2.14 (m, 1H), 1.99-1.94 (m, 2H),1.83-1.75 (m, 2H), 1.51-1.38 (m, 1H), 1.32-1.22 (m, 1H), 0.86 (d, J=6.0Hz, 3H).

¹⁹F NMR (400 MHz, CD₃OD): δ −69.39.

Elemental Analysis: Calcd for C₃₁H₄₃F₃N₄O₈. H₂O: C, 55.18; H, 6.72; N,8.30. Found: C, 54.95; H, 6.89; N, 8.07.

Moisture Content (Karl Fischer): 2.33%

The elemental analysis of Compound B meglumine salt shows that itcontains a one to one to ratio of Compound A cation to meglumine anionto water molecule.

1.-9. (canceled)
 10. A compound of Formula (II):

in the form of a meglumine salt or a hydrate of the meglumine salt. 11.The compound of claim 10, wherein the compound is in the form of amonohydrate.
 12. The compound of claim 10, wherein the compound isunhydrated.
 13. A pharmaceutical composition comprising the compound ofclaim 10, and a pharmaceutically acceptable excipient.
 14. A method oftreating a PPARδ related disease or condition in a subject, comprisingadministering to the subject in need thereof a therapeutically effectiveamount of the compound of claim
 10. 15. The method of claim 14, whereinthe PPARδ related disease is a muscle structure disorder, a neuronalactivation disorder, a muscle fatigue disorder, a muscle mass disorder,a mitochondrial disease, a beta oxidation disease, a metabolic disease,a cancer, a vascular disease, an ocular vascular disease, a muscular eyedisease, or a renal disease.
 16. The method of claim 15, wherein: themuscle structure disorder is selected from Bethlem myopathy, centralcore disease, congenital fiber type disproportion, distal musculardystrophy (MD), Duchenne & Becker MD, Emery-Dreifuss MD,facioscapulohumeral MD, hyaline body myopathy, limb-girdle MD, a musclesodium channel disorders, myotonic chondrodystrophy, myotonic dystrophy,myotubular myopathy, nemaline body disease, oculopharyngeal MD, orstress urinary incontinence; the neuronal activation disorder isselected from amyotrophic lateral sclerosis, Charcot-Marie-Toothdisease, Guillain-Barre syndrome, Lambert-Eaton syndrome, multiplesclerosis, myasthenia gravis, nerve lesion, peripheral neuropathy,spinal muscular atrophy, tardy ulnar nerve palsy, or toxic myoneuraldisorder; the muscle fatigue disorder is selected from chronic fatiguesyndrome, diabetes (type I or II), glycogen storage disease,fibromyalgia, Friedreich's ataxia, intermittent claudication, lipidstorage myopathy, MELAS, mucopolysaccharidosis, Pompe disease, orthyrotoxic myopathy; the muscle mass disorder is cachexia, cartilagedegeneration, cerebral palsy, compartment syndrome, critical illnessmyopathy, inclusion body myositis, muscular atrophy (disuse),sarcopenia, steroid myopathy, or systemic lupus erythematosus; themitochondrial disease is selected from Alpers's Disease, CPEO-Chronicprogressive external ophthalmoplegia, Kearns-Sayra Syndrome (KSS), LeberHereditary Optic Neuropathy (LHON), MELAS-Mitochondrial myopathy,encephalomyopathy, lactic acidosis, and stroke-like episodes,MERRF-Myoclonic epilepsy and ragged-red fiber disease, NARP-neurogenicmuscle weakness, ataxia, and retinitis pigmentosa, or Pearson Syndrome;the beta oxidation disease is selected from systemic carnitinetransporter, carnitine palmitoyltransferase (CPT) II deficiency, verylong-chain acyl-CoA dehydrogenase (LCHAD or VLCAD) deficiency,trifunctional enzyme deficiency, medium-chain acyl-CoA dehydrogenase(MCAD) deficiency, short-chain acyl-CoA dehydrogenase (SCAD) deficiencyor riboflavin-responsive disorders of β-oxidation (RR-MADD); themetabolic disease is selected from hyperlipidemia, dyslipidemia,hyperchlolesterolemia, hypertriglyceridemia, HDL hypocholesterolemia,LDL hypercholesterolemia and/or HLD non-cholesterolemia, VLDLhyperproteinemia, dyslipoproteinemia, apolipoprotein A-Ihypoproteinemia, atherosclerosis, disease of arterial sclerosis, diseaseof cardiovascular systems, cerebrovascular disease, peripheralcirculatory disease, metabolic syndrome, syndrome X, obesity, diabetes(type I or II), hyperglycemia, insulin resistance, impaired glucosetolerance, hyperinsulinism, diabetic complication, cardiacinsufficiency, cardiac infarction, cardiomyopathy, hypertension,Non-alcoholic fatty liver disease (NAFLD), Nonalcoholic steatohepatitis(NASH), thrombus, Alzheimer disease, neurodegenerative disease,demyelinating disease, multiple sclerosis, adrenoleukodystrophy,dermatitis, psoriasis, acne, skin aging, trichosis, inflammation,arthritis, asthma, hypersensitive intestine syndrome, ulcerativecolitis, Crohn's disease, or pancreatitis; the cancer is a cancer of thecolon, large intestine, skin, breast, prostate, ovary, or lung; thevascular disease is selected from peripheral vascular insufficiency,peripheral vascular disease, intermittent claudication, peripheralvascular disease (PVD), peripheral artery disease (PAD), peripheralartery occlusive disease (PAOD), or peripheral obliterativearteriopathy; the ocular vascular disease is selected from age-relatedmacular degeneration (AMD), stargardt disease, hypertensive retinopathy,diabetic retinopathy, retinopathy, macular degeneration, retinalhaemorrhage, or glaucoma; the muscular eye disease is selected fromstrabismus, progressive external ophthalmoplegia, esotropia, exotropia,a disorder of refraction and accommodation, hypermetropia, myopia,astigmatism, anisometropia, presbyopia, a disorders of accommodation, orinternal ophthalmoplegia; and the renal disease is selected fromglomerulonephritis, glomerulosclerosis, nephrotic syndrome, hypertensivenephrosclerosis, acute nephritis, recurrent hematuria, persistenthematuria, chronic nephritis, rapidly progressive nephritis, acutekidney injury, chronic renal failure, diabetic nephropathy, or Bartter'ssyndrome.
 17. A method of treating acute kidney injury in a subject,comprising administering to the subject in need thereof atherapeutically effective amount of a compound of claim
 10. 18. A methodof treating a mitochondrial disease in a subject, comprisingadministering to the subject in need thereof a therapeutically effectiveamount of a compound of claim 10, wherein the mitochondrial disease isAlpers Disease, chronic progressive external ophthalmoplegia (CPEO),Kearns-Sayra Syndrome (KSS), Leber Hereditary Optic Neuropathy (LHON),MELAS-mitochondrial myopathy, encephalomyopathy, lactic acidosis, andstroke-like episodes, myoclonic epilepsy with ragged-red fiber disease(MERRF), neurogenic muscle weakness-ataxia-retinitis pigmentosa (NARP),or Pearson Syndrome.